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Chemogenetic Regulation in Reprogrammed Stem Cell-derived Precursor Cells in Treating Neurodegenerative Diseases

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机构: [1]Cell Therapy Center, Beijing Municipal Geriatric Medical Research Center, Xuanwu Hospital Capital Medical University [2]National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education,Xuanwu Hospital Capital Medical University. [3]The Department of Neurology, Xuanwu Hospital Capital Medical University.
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DOI: 10.3791/68032

摘要:
The integration of designer receptors exclusively activated by designer drugs (DREADD) with stem cell-based therapies presents an advanced strategy for precise neuronal modulation. Here, we utilized CRISPR-engineered human reprogrammed stem cells expressing excitatory (hM3Dq) or inhibitory (hM4Di) DREADD receptors to evaluate the functional integration and modulation of transplanted dopaminergic precursors in a murine model of Parkinson's disease (PD). Key steps included generating non-fusion DREADD-expressing stem cell lines, differentiating them into midbrain dopaminergic precursors, and transplanting these cells into the striatum of 6-hydroxydopamine (6-OHDA)-lesioned mice. We conducted behavioral assessments and electrophysiological recordings to analyze the effects of the transplanted cells. Behavioral tests, such as the cylinder test, demonstrated significant modulation of motor function following clozapine-N-oxide (CNO) administration. Specifically, activation of hM4Di reduced contralateral forelimb movement, whereas activation of hM3Dq was associated with enhanced motor behavior. Electrophysiological recordings revealed distinct synaptic responses. hM4Di activation increased interevent intervals and decreased peak amplitudes of spontaneous excitatory postsynaptic currents (sEPSCs), whereas hM3Dq activation decreased interevent intervals and increased peak amplitudes, reflecting enhanced excitatory signaling. In summary, the integration of behavioral and electrophysiological assessments validates the precise functional incorporation of engineered chemically reprogrammed stem cells into host neural circuits.

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大类 | 4 区 综合性期刊
小类 | 4 区 综合性期刊
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大类 | 4 区 综合性期刊
小类 | 4 区 综合性期刊
第一作者:
第一作者机构: [1]Cell Therapy Center, Beijing Municipal Geriatric Medical Research Center, Xuanwu Hospital Capital Medical University [2]National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education,Xuanwu Hospital Capital Medical University.
通讯作者:
通讯机构: [1]Cell Therapy Center, Beijing Municipal Geriatric Medical Research Center, Xuanwu Hospital Capital Medical University [2]National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education,Xuanwu Hospital Capital Medical University.
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